小鼠口服猪卵透明带DNA避孕疫苗pVAX1-pZP3α的抗生育研究  

Anti-fertility study of oral immunization in mouse with DNA contraceptive vaccine Chitosan/pVAX1-pZP3α nanoparticles

作　　者：董志炜[1] 潘善培[1] 谢琪璇[1] 肖銮娟[1] 孙彩军[1] 余巍[1] 何柳媚[1] 

机构地区：[1]暨南大学生殖免疫研究所,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2004年第6期661-667,共7页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金资助项目(30370541和30230350)

摘　　要：　目的:评价口服卵透明带DNA避孕疫苗pVAX1-pZP3α壳聚糖纳米微粒诱发的黏膜免疫反应、抗生育作用和对卵巢结构的影响,探讨用黏膜免疫避免卵透明带抗原免疫引起卵巢功能紊乱的可行性。方法:制备pVAX1-pZP3α壳聚糖纳米微粒,10只昆明系小鼠口服免疫,第3周和第6周分别加强免疫一次,剂量为0 5mL/只,含pVAX1-pZP3αDNA50μg;对照组10只雌小鼠口服相同剂量不含pVAX1-pZP3α的壳聚糖。免疫前和免疫后隔周取小鼠血清、阴道冲洗液,用ELISA法检测抗pZP3IgA和IgG。首次免疫后第12周雌雄合笼交配,统计雌鼠怀孕情况;解剖取出双侧卵巢,制作石蜡切片,光镜观察卵巢病理学变化。结果:免疫组80%小鼠的血清和50%小鼠的阴道冲洗液可检测到抗pZP3IgA,反应强度与持续时间有明显的个体差异;免疫组5只小鼠未怀孕,对照组全部怀孕,抗生育效果与合笼时抗pZP3IgA反应有关;免疫组未怀孕小鼠卵巢组织结构与免疫组怀孕小鼠及对照组小鼠无差别。结论:口服pVAX1-pZP3α壳聚糖纳米微粒免疫小鼠能诱发生殖道黏膜免疫反应,发挥抗生育作用,对卵巢组织结构没有影响。黏膜免疫是克服卵透明带抗原免疫引起卵巢功能紊乱的一种值得深入研究的方法。Aim: To evaluate the mucosal immune response, anti-fertility, change of ovarian histology induced by oral immunization with DNA contraceptive vaccine Chitosan/pVAX1-pZP3α nanoparticles and explore the feasibility of taking advantage of mucosal immunity to overcome ovary dysfunction which was often caused by immunization of zona pellucida antigen. Methods: Female mice were divided into two different groups of ten. 0.5 mL Chitosan/pVAX1-pZP3α nanoparticles containing 50 μg DNA was prepared and orally administered into each experimental mouse on weeks 0,3 and 6 whereas the same dose of chitosan without DNA for the control. Serum and vaginal wash samples collected every two weeks were used to determine anti-pZP3 IgA and IgG response by ELISA. On week 12, these female mice were mated with male mice and were checked for pregnant. On week 16, the mice were killed and ovarian serial section was prepared for microscopy. Results: Anti-pZP3 IgA antibody response could be detected in serum from eight experimental mice and vaginal washes from five, intensity and lasting time varying over a wide range among different mouse. Five experimental mice didn't fall pregnant while the control did and anti-fertility was relative to Anti-pZP3 IgA antibody level in mating mice. There was no difference about ovarian histology among unpregnant, pregnant experimental mice and control mice. Conclusion:Oral immunization with chitosan/pVAX1-pZP3α nanoparticles could induce mucosal immune response in genital tract and inhibit fertility without any effect on ovarian structure in mice. Inducing mucosal immunity would be a promising regimen for study of developing a safe zona pellucida contraceptive vaccine.

关 键 词：卵透明带 避孕疫苗 黏膜免疫 DNA疫苗 卵巢功能紊乱 

分 类 号：Q784[生物学—分子生物学] R16941[医药卫生—公共卫生与预防医学]